Identification of a plasminogen binding region in streptokinase that is necessary for the creation of a functional streptokinase-plasminogen activator complex
Streptokinase is a plasminogen activator widely used to treat patients with myocardial infarction. However, streptokinase is not a protease, and must first bind and interact with plasminogen to form an enzymatic complex. By measuring the binding of recombinant streptokinase fragments to plasminogen,...
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Veröffentlicht in: | Biochemistry (Easton) 1995-08, Vol.34 (32), p.10266-10271 |
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creator | Reed, Guy L Lin, Lee-Fong Parhami-Seren, Behnaz Kussie, Paul |
description | Streptokinase is a plasminogen activator widely used to treat patients with myocardial infarction. However, streptokinase is not a protease, and must first bind and interact with plasminogen to form an enzymatic complex. By measuring the binding of recombinant streptokinase fragments to plasminogen, we have sought, first, to identify a plasminogen binding region in streptokinase and, second, to explore the relation between binding (via this region) and the generation of a functional streptokinase--plasminogen activator complex. Recombinant streptokinase bound in a saturable and specific manner to human Glu-plasminogen with a dissociation constant of 4.2 x 10(-10) M. Recombinant streptokinase fragments spanning amino acids 1-127 and 1-253 could not be shown to bind to Glu-plasminogen, whereas fragments spanning amino acids 1-352, 120-352, and 244-414 bound tightly to plasminogen and each fragment completely inhibited the binding of full-length streptokinase to plasminogen. Although these latter streptokinase fragments formed a complex with plasminogen, enzymatic assays indicated that none of them was capable of generating an active site. When the streptokinase region shared by these three fragments, spanning residues 244-352, was expressed, it also bound plasminogen and competitively inhibited the formation of a functional plasminogen activator complex by full-length streptokinase. Taken together, these data indicate that streptokinase binds to plasminogen with high affinity, that a primary binding region for plasminogen is located within amino acids 244-352, and that binding via this region is necessary for the generation of a functional plasminogen activator complex. |
doi_str_mv | 10.1021/bi00032a021 |
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However, streptokinase is not a protease, and must first bind and interact with plasminogen to form an enzymatic complex. By measuring the binding of recombinant streptokinase fragments to plasminogen, we have sought, first, to identify a plasminogen binding region in streptokinase and, second, to explore the relation between binding (via this region) and the generation of a functional streptokinase--plasminogen activator complex. Recombinant streptokinase bound in a saturable and specific manner to human Glu-plasminogen with a dissociation constant of 4.2 x 10(-10) M. Recombinant streptokinase fragments spanning amino acids 1-127 and 1-253 could not be shown to bind to Glu-plasminogen, whereas fragments spanning amino acids 1-352, 120-352, and 244-414 bound tightly to plasminogen and each fragment completely inhibited the binding of full-length streptokinase to plasminogen. Although these latter streptokinase fragments formed a complex with plasminogen, enzymatic assays indicated that none of them was capable of generating an active site. When the streptokinase region shared by these three fragments, spanning residues 244-352, was expressed, it also bound plasminogen and competitively inhibited the formation of a functional plasminogen activator complex by full-length streptokinase. Taken together, these data indicate that streptokinase binds to plasminogen with high affinity, that a primary binding region for plasminogen is located within amino acids 244-352, and that binding via this region is necessary for the generation of a functional plasminogen activator complex.</description><identifier>ISSN: 0006-2960</identifier><identifier>EISSN: 1520-4995</identifier><identifier>DOI: 10.1021/bi00032a021</identifier><identifier>PMID: 7640282</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Base Sequence ; Binding Sites ; Cloning, Molecular ; Molecular Sequence Data ; Oligodeoxyribonucleotides ; Peptide Fragments - chemistry ; Peptide Fragments - metabolism ; Plasminogen - metabolism ; Plasminogen Activators - chemistry ; Plasminogen Activators - metabolism ; Streptokinase - chemistry ; Streptokinase - genetics ; Streptokinase - metabolism</subject><ispartof>Biochemistry (Easton), 1995-08, Vol.34 (32), p.10266-10271</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a354t-7d41bcb43c052ffa6138d634580ca2fbad4f507561de6627f8693edc1bd23bb13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://pubs.acs.org/doi/pdf/10.1021/bi00032a021$$EPDF$$P50$$Gacs$$H</linktopdf><linktohtml>$$Uhttps://pubs.acs.org/doi/10.1021/bi00032a021$$EHTML$$P50$$Gacs$$H</linktohtml><link.rule.ids>314,780,784,2765,27076,27924,27925,56738,56788</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7640282$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Reed, Guy L</creatorcontrib><creatorcontrib>Lin, Lee-Fong</creatorcontrib><creatorcontrib>Parhami-Seren, Behnaz</creatorcontrib><creatorcontrib>Kussie, Paul</creatorcontrib><title>Identification of a plasminogen binding region in streptokinase that is necessary for the creation of a functional streptokinase-plasminogen activator complex</title><title>Biochemistry (Easton)</title><addtitle>Biochemistry</addtitle><description>Streptokinase is a plasminogen activator widely used to treat patients with myocardial infarction. However, streptokinase is not a protease, and must first bind and interact with plasminogen to form an enzymatic complex. By measuring the binding of recombinant streptokinase fragments to plasminogen, we have sought, first, to identify a plasminogen binding region in streptokinase and, second, to explore the relation between binding (via this region) and the generation of a functional streptokinase--plasminogen activator complex. Recombinant streptokinase bound in a saturable and specific manner to human Glu-plasminogen with a dissociation constant of 4.2 x 10(-10) M. Recombinant streptokinase fragments spanning amino acids 1-127 and 1-253 could not be shown to bind to Glu-plasminogen, whereas fragments spanning amino acids 1-352, 120-352, and 244-414 bound tightly to plasminogen and each fragment completely inhibited the binding of full-length streptokinase to plasminogen. Although these latter streptokinase fragments formed a complex with plasminogen, enzymatic assays indicated that none of them was capable of generating an active site. When the streptokinase region shared by these three fragments, spanning residues 244-352, was expressed, it also bound plasminogen and competitively inhibited the formation of a functional plasminogen activator complex by full-length streptokinase. Taken together, these data indicate that streptokinase binds to plasminogen with high affinity, that a primary binding region for plasminogen is located within amino acids 244-352, and that binding via this region is necessary for the generation of a functional plasminogen activator complex.</description><subject>Base Sequence</subject><subject>Binding Sites</subject><subject>Cloning, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Oligodeoxyribonucleotides</subject><subject>Peptide Fragments - chemistry</subject><subject>Peptide Fragments - metabolism</subject><subject>Plasminogen - metabolism</subject><subject>Plasminogen Activators - chemistry</subject><subject>Plasminogen Activators - metabolism</subject><subject>Streptokinase - chemistry</subject><subject>Streptokinase - genetics</subject><subject>Streptokinase - metabolism</subject><issn>0006-2960</issn><issn>1520-4995</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1995</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkUFv1DAQhS0EKkvhxBnJJziggO04dnJEBdpKlUC0nC3bGS9uEzvYDip_ht-KV7uqFomTPfM-vaeZQeglJe8oYfS98YSQlun6f4Q2tGOk4cPQPUab2hcNGwR5ip7lfFtLTiQ_QSdScMJ6tkF_LkcIxTtvdfEx4Oiwxsuk8-xD3ELAxofRhy1OsN3pPuBcEiwl3vmgM-DyQxfsMw5gIWedfmMXU-0CtgmOPN0a7K7S078GzXGYrsgvXaqBjfMywf1z9MTpKcOLw3uKvn_-dHN20Vx9Ob88-3DV6LbjpZEjp8Ya3lrSMee0oG0_ipZ3PbGaOaNH7joiO0FHEIJJ14uhhdFSM7LWGNqeotd73yXFnyvkomafLUyTDhDXrKTknMuhr-DbPWhTzDmBU0vycx1bUaJ211BH16j0q4PtamYYH9jD-qve7HWfC9w_yDrdKSFb2ambr9eKfPt4fX4hidqlv9nz2mZ1G9dU15n_m_wX5w2k3A</recordid><startdate>19950815</startdate><enddate>19950815</enddate><creator>Reed, Guy L</creator><creator>Lin, Lee-Fong</creator><creator>Parhami-Seren, Behnaz</creator><creator>Kussie, Paul</creator><general>American Chemical Society</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>19950815</creationdate><title>Identification of a plasminogen binding region in streptokinase that is necessary for the creation of a functional streptokinase-plasminogen activator complex</title><author>Reed, Guy L ; Lin, Lee-Fong ; Parhami-Seren, Behnaz ; Kussie, Paul</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a354t-7d41bcb43c052ffa6138d634580ca2fbad4f507561de6627f8693edc1bd23bb13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1995</creationdate><topic>Base Sequence</topic><topic>Binding Sites</topic><topic>Cloning, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Oligodeoxyribonucleotides</topic><topic>Peptide Fragments - chemistry</topic><topic>Peptide Fragments - metabolism</topic><topic>Plasminogen - metabolism</topic><topic>Plasminogen Activators - chemistry</topic><topic>Plasminogen Activators - metabolism</topic><topic>Streptokinase - chemistry</topic><topic>Streptokinase - genetics</topic><topic>Streptokinase - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Reed, Guy L</creatorcontrib><creatorcontrib>Lin, Lee-Fong</creatorcontrib><creatorcontrib>Parhami-Seren, Behnaz</creatorcontrib><creatorcontrib>Kussie, Paul</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Biochemistry (Easton)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Reed, Guy L</au><au>Lin, Lee-Fong</au><au>Parhami-Seren, Behnaz</au><au>Kussie, Paul</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Identification of a plasminogen binding region in streptokinase that is necessary for the creation of a functional streptokinase-plasminogen activator complex</atitle><jtitle>Biochemistry (Easton)</jtitle><addtitle>Biochemistry</addtitle><date>1995-08-15</date><risdate>1995</risdate><volume>34</volume><issue>32</issue><spage>10266</spage><epage>10271</epage><pages>10266-10271</pages><issn>0006-2960</issn><eissn>1520-4995</eissn><abstract>Streptokinase is a plasminogen activator widely used to treat patients with myocardial infarction. However, streptokinase is not a protease, and must first bind and interact with plasminogen to form an enzymatic complex. By measuring the binding of recombinant streptokinase fragments to plasminogen, we have sought, first, to identify a plasminogen binding region in streptokinase and, second, to explore the relation between binding (via this region) and the generation of a functional streptokinase--plasminogen activator complex. Recombinant streptokinase bound in a saturable and specific manner to human Glu-plasminogen with a dissociation constant of 4.2 x 10(-10) M. Recombinant streptokinase fragments spanning amino acids 1-127 and 1-253 could not be shown to bind to Glu-plasminogen, whereas fragments spanning amino acids 1-352, 120-352, and 244-414 bound tightly to plasminogen and each fragment completely inhibited the binding of full-length streptokinase to plasminogen. Although these latter streptokinase fragments formed a complex with plasminogen, enzymatic assays indicated that none of them was capable of generating an active site. When the streptokinase region shared by these three fragments, spanning residues 244-352, was expressed, it also bound plasminogen and competitively inhibited the formation of a functional plasminogen activator complex by full-length streptokinase. Taken together, these data indicate that streptokinase binds to plasminogen with high affinity, that a primary binding region for plasminogen is located within amino acids 244-352, and that binding via this region is necessary for the generation of a functional plasminogen activator complex.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>7640282</pmid><doi>10.1021/bi00032a021</doi><tpages>6</tpages></addata></record> |
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subjects | Base Sequence Binding Sites Cloning, Molecular Molecular Sequence Data Oligodeoxyribonucleotides Peptide Fragments - chemistry Peptide Fragments - metabolism Plasminogen - metabolism Plasminogen Activators - chemistry Plasminogen Activators - metabolism Streptokinase - chemistry Streptokinase - genetics Streptokinase - metabolism |
title | Identification of a plasminogen binding region in streptokinase that is necessary for the creation of a functional streptokinase-plasminogen activator complex |
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